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DNA double-strand breaks repair inhibitors potentiates the combined effect of VP-16 and CDDP in human colorectal adenocarcinoma (LoVo) cells

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Abstract

I. Background: A combination of etoposide (VP-16) and cisplatin (CDDP) is the standard treatment for certain colon cancers. These drugs promote the death of cancer cells via direct and indirect induction of the most lethal DNA lesions – DNA double-stand breaks. However, cancer cells can reverse the DNA damaging effect of anticancer drugs by triggering DNA repair processes. In eukaryotic cells, the main DNA repair pathway responsible for DNA double-stand breaks repair is non-homologous end-joining (NHEJ). Inhibitors of DNA repair are of special interest in cancer research as they could break the cellular resistance to DNA-damaging agents and increase the efficiency of standard cancer treatments. In this study, we investigated the effect of two NHEJ inhibitors, SCR7 and NU7441, on the cytotoxic mechanism of VP-16/CDDP in a LoVo human colorectal adenocarcinoma cell line. SCR7 blocks Ligase IV-mediated joining by interfering with its DNA binding, whereas NU7441 is a highly potent and selective DNA-PK inhibitor.

II. Methods and Results: Both inhibitors synergistically increased the cytotoxicity of CDDP and VP-16 when combined, but the effect of SCR7 was more pronounced. SCR7 and NU7441 also significantly increased VP-16; CDDP induced DNA double-stand breaks level and delayed drug-induced DSB repair, as seen on the comet assay and measured using H2AX foci. We also observed changes in cell cycle distribution and enhanced apoptosis ratio in colorectal adenocarcinoma cells treated with DNA repair inhibitors and VP-16/CDDP.

III. Conclusions: Our data support the hypothesis that NHEJ inhibitors could be used in conjunction with standard therapy to provide effective clinical improvement and allow reduction in drug doses.

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Availability of data and material

Raw data were generated at University of Lodz. Derived data supporting the findings of this study are available from the corresponding author TP on request.

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Acknowledgements

This work was financially supported by the National Science Centre (NCN, Poland) grant, according to the decision No. DEC-2013/11/B/NZ7/01340.

Funding

This work was financially supported by the National Science Center (NCN, Poland) grant, according to the decision No. DEC-2013/11/B/NZ7/01340.

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Authors and Affiliations

  1. Faculty of Medicine, Department of Immunopathology, Division of Biomedical Science, Medical University of Lodz, Lodz, Poland

    Paulina Kopa

  2. Department of Molecular Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland

    Anna Macieja & Tomasz Poplawski

  3. Functional Genomics & Proteomics Unit, ITSI-Biosciences, Johnstown, PA, USA

    Elzbieta Pastwa

  4. Department of Clinical Chemistry and Biochemistry, Faculty of Medicine, Medical University of Lodz, Lodz, Poland

    Ireneusz Majsterek

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  1. Paulina Kopa
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  4. Ireneusz Majsterek
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  5. Tomasz Poplawski
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Contributions

PK performed the experiments, analyzed the data, interpreted the results and wrote the manuscript; AM participated in the laboratory work, interpreting results and wrote the manuscript; EP and IM participated in the design of the study, critically reviewed the manuscript; TP supervised overall study, analyzed data, and critically reviewed manuscript. All authors read and approved the final manuscript.

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Correspondence to Tomasz Poplawski.

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Kopa, P., Macieja, A., Pastwa, E. et al. DNA double-strand breaks repair inhibitors potentiates the combined effect of VP-16 and CDDP in human colorectal adenocarcinoma (LoVo) cells. Mol Biol Rep 48, 709–720 (2021). https://doi.org/10.1007/s11033-020-06124-9

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